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Quantum defects as versatile anchors for carbon nanotube functionalization

Nature Protocols volume 17, pages 727–747 (2022)Cite this article

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Abstract

Single-wall carbon nanotubes (SWCNTs) are used in diverse applications that require chemical tailoring of the SWCNT surface, including optical sensing, imaging, targeted drug delivery and single-photon generation. SWCNTs have been noncovalently modified with (bio)polymers to preserve their intrinsic near-infrared fluorescence. However, demanding applications (e.g., requiring stability in biological fluids) would benefit from a stable covalent linkage between the SWCNT and the functional unit (e.g., antibody, fluorophore, drug). Here we present how to use diazonium salt chemistry to introduce sp3 quantum defects in the SWCNT carbon lattice to serve as handles for conjugation while preserving near-infrared fluorescence. In this protocol, we describe the straightforward, stable (covalent), highly versatile and scalable functionalization of SWCNTs with biomolecules such as peptides and proteins to yield near-infrared fluorescent SWCNT bioconjugates. We provide a step-by-step procedure covering SWCNT dispersion, quantum defect incorporation, bioconjugation, in situ peptide synthesis on SWCNTs, and characterization, which can be completed in 5–7 d.

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Fig. 1: Modular chemistry with quantum defects on SWCNTs.
Fig. 2: Workflow of quantum defect incorporation and bioconjugation.
Fig. 3: Custom-built NIR fluorescence spectrometer.
Fig. 4: Well-plate system for light-catalyzed reactions.
Fig. 5: Observation of the defect reaction by fluorescence spectroscopy.
Fig. 6: Larger-scale defect incorporation.
Fig. 7: SWCNT–peptide synthesis in a 96-well filter plate.
Fig. 8: Assessment of successful functionalization.
Fig. 9: Assessment of SWCNT–peptide synthesis.

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Data availability

The authors declare that the main data discussed in this protocol are available in the supporting primary research paper35. The raw datasets are available for research purposes from the corresponding authors upon reasonable request and are publicly available in the repository https://doi.org/10.6084/m9.figshare.16595093.

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Acknowledgements

This work was funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) under Germany’s Excellence Strategy–EXC 2033–390677874–RESOLV. This project was supported by the VW Foundation.

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Authors and Affiliations

  1. Physical Chemistry II, Ruhr-Universität Bochum, Bochum, Germany

    Florian A. Mann, Phillip Galonska & Sebastian Kruss

  2. Institute of Physical Chemistry, Göttingen University, Göttingen, Germany

    Florian A. Mann, Niklas Herrmann & Sebastian Kruss

  3. Fraunhofer Institute for Microelectronic Circuits and Systems, Duisburg, Germany

    Sebastian Kruss

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Contributions

S.K. and F.A.M. designed and conceived the project. S.K. coordinated the project. N.H. performed MalPh diazonium salt synthesis, NMR/MS analysis and initial experiments, and optimized the conditions for MalPh-defect incorporation. F.A.M. performed AFM experiments, synthesized the Fmoc-Phe diazonium salt with N.H., optimized the conditions for Fmoc-Phe defect incorporation, performed the nanobody conjugation and validation experiments, synthesized multicolor SWCNTs, SWCNT-R6 and performed the 96-well SWCNT–peptide synthesis as well as the subsequent characterization. F.A.M., P.G., N.H. and S.K wrote the manuscript.

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Correspondence to Sebastian Kruss.

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Nature Protocols thanks Hyejin Kwon and YuHuang Wang for their contribution to the peer review of this work.

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Mann, F. A. et al. Angew. Chem. Int. Ed. Engl. 59,17732–17738 (2020): https://doi.org/10.1002/anie.202003825

Spreinat, A. et al. J. Phys. Chem. C 125, 18341–18351 (2021): https://doi.org/10.1021/acs.jpcc.1c05432

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Mann, F.A., Galonska, P., Herrmann, N. et al. Quantum defects as versatile anchors for carbon nanotube functionalization. Nat Protoc 17, 727–747 (2022). https://doi.org/10.1038/s41596-021-00663-6

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